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Generation of Growth Curve

Adapted from Mather, J.P., and P.E. Roberts, 1998. Introduction to Cell and Tissue Culture: Theory and Technique. Plenum Press. New York and London.

Generation of a growth curve can be useful in evaluating the growth characteristics of a cell line. From a growth curve, the lag time, population doubling time, and saturation density can be determined.

Materials


1.

Cells
2. Tissue culture dishes or flasks
3. Growth medium
4. Trypsin

 

 

Procedure


1.

Trypsinize the cells (see Passage of Adherent Cell Lines) and centrifuge the cells.
2. Resuspend the pellet in 5 mL of medium and count the cells (see Hemocytometer Counting.)
3. Dilute the cell suspension in order to have an appropriate amount of medium and cells to achieve a seeding density of 2 x 103 cells per cm2 of surface area. Cell seeding densities vary by cell line.
4. Mix well and seed the dishes/flasks with the appropriate amount of diluted cell suspension.
5.

Count some of the leftover cell suspension in order to determine the actual seeding density.
6. Put the Plates in an incubator.
7. Count the duplicate plates every 24 hours.
8. Plot the results on a log-linear scale. The population-doubling time can be determined by identifying a cell number along the exponential phase of the curve, tracing the curve until that number has doubled, and calculating the time between the two.

 

 

 

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